Journal of Oral Microbiology | |
Metatranscriptomics reveals overall active bacterial composition in caries lesions | |
Alex Mira2  Áurea Simón-Soro1  Miriam Guillén-Navarro1  | |
[1] FISABIO Foundation, Center for Advanced Research in Public Health;FISABIO Foundation, Center for Advanced Research in Public Health | |
关键词: 16S rRNA; pyrosequencing; Streptoccus mutans; polymicrobial disease; tissue-dependent hypothesis; caries etiology; | |
DOI : | |
来源: Co-Action Publishing | |
【 摘 要 】
Background:Identifyingthemicrobialspeciesincarieslesionsisinstrumentaltodeterminetheetiologyofdentalcaries.However,asignificantproportionofbacteriaincariouslesionshavenotbeencultured,andtheuseofmolecularmethodshasbeenlimitedtoDNA-basedapproaches,whichdetectbothactiveandinactiveordeadmicroorganisms.Objective:ToidentifytheRNA-based,metabolicallyactivebacterialcompositionofcarieslesionsatdifferentstagesofdiseaseprogressioninordertoprovidealistofpotentialetiologicalagentsoftoothdecay.Design:Non-cavitatedenamelcarieslesions(n=15)anddentincarieslesionssamples(n=12)werecollectedfrom13individuals.RNAwasextractedandcDNAwasconstructed,whichwasusedtoamplifythe16SrRNAgene.Theresulting780bppolymerasechainreactionproductswerepyrosequencedusingTitanium-pluschemistry,andthesequencesobtainedwereusedtodeterminethebacterialcomposition.Results:Ameanof4,900sequencesofthe16SrRNAgenewithanaveragereadlengthof661bpwasobtainedpersample,givingacomprehensiveviewoftheactivebacterialcommunitiesincarieslesions.Estimatesofbacterialdiversityindicatethatthemicrobiotaofcavitiesishighlycomplex,eachsamplecontainingbetween70and400metabolicallyactivespecies.Thecompositionofthesebacterialconsortiavariedamongindividualsandbetweencarieslesionsofthesameindividuals.Inaddition,enamelanddentinlesionshadadifferentbacterialmakeup.Lactobacilliwerefoundalmostexclusivelyindentincavities.Streptococciaccountedfor40%ofthetotalactivecommunityinenamelcaries,and20%indentincaries.However,Streptococcusmutansrepresentedonly0.02–0.73%ofthetotalbacterialcommunity.Conclusions:Thedataindicatethattheetiologyofdentalcariesistissuedependentandthatthediseasehasaclearpolymicrobialorigin.Thelowproportionofmutansstreptococcidetectedconfirmsthattheyareaminorityandquestionsitsimportanceasthemainetiologicalagentoftoothdecay.Futureexperimentalworkshouldbeperformedtoconfirmthecariogenicityoftheidentifiedbacteria.Keywords:16SrRNA;pyrosequencing;Streptococcusmutans;polymicrobialdisease;tissue-dependenthypothesis;cariesetiology(Published:24October2014)Citation:JournalofOralMicrobiology2014,6:25443-http://dx.doi.org/10.3402/jom.v6.25443
【 授权许可】
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